Fig. 1

Effects of Ventromedial Hypothalamic Nucleus (VMN) Neuronal Nitric Oxide Synthase (nNOS) Gene Silencing on Dorsomedial VMN (VMNdm) or Ventrolateral VMN (VMNvl) γ-Aminobutyric Acid (GABA) Neuron nNOS Gene Expression in the Male Rat. Groups of adult male rats (n = 6/group) were pretreated by bilateral stereotactic administration of control/scramble (SCR) or nNOS siRNA to the VMN, then injected subcutaneously (sc) with vehicle (V) or neutral protamine Hagedorn insulin (INS; 10.0 U/kg bw) one hour before sacrifice. Individual VMNdm and VMNvl GABAergic neurons were labeled in situ by immunocytochemical methods for GAD65/67-immunoreactivity (-ir) prior to laser-catapult-microdissection for single-cell multiplex qPCR analysis. mRNA data were normalized to the housekeeping gene GAPDH by the 2^−ΔΔCt method. Gene expression data are presented here in box-and-whisker plot format, which displays the median, lower and upper quartiles, and lower and upper extremes of a data set. Plots depict normalized nNOS values for VMNdm (Fig. 1A) and VMNvl Fig. 1B) GABAergic neurons collected from the following treatment groups: SCR siRNA/sc V (blue box-and-whisker plots, n = 12), nNOS siRNA/sc V (yellow box-and-whisker plots; n = 12), SCR siRNA/ sc INS (green box-and-whisker plots; n = 12), nNOS siRNA/sc INS (orange box-and-whisker plots; n = 12). Statistical differences between discrete pairs of treatment groups are denoted as follows: *p < 0.05; **p < 0.01; ***p < 0.001